Multiple births represent a significant cause of childhood morbidity and long-term compromised health. In vitro fertilization (IVF) treatment has significantly increased the number of multiple births by the transfer multiple embryos in a single cycle. Current morphological criteria are not sufficient to precisely identify those embryos that will implant successfully. Our preliminary studies based on proton nuclear magnetic resonance spectroscopy (1H-NMR) show that multivariate analysis of the NMR spectra recorded for culture media samples from 100% and 0% implantation cases revealed a statistical separation of the two groups (p=0.009). In addition, deconvolution of the NMR spectra into the contributions from individual compounds present in the culture media suggested that the concentrations of several compounds were significantly different between the 100% and 0% implantation classes. Based on these comprehensive preliminary studies, we thus hypothesize that implantable embryos have metabolic characteristics that can be identified in conditioned culture media using 1D 1H-NMR. To test our hypothesis, we propose to establish NMR-based profiling of spent culture media as a tool to non-invasively predict embryo implantation success. In Specific Aim 1 we will develop statistical models to predict embryo implantation success based on 1D 1H NMR spectra recorded for media from 0% versus 100% implantation cases. In Specific Aim 2, we will test the robustness of these predictive models by applying our protocol for NMR-based profiling to new media specimens from cases with partial embryo implantation success. This will enable us to evaluate the models'sensitivity, specificity, and predictive value. The significance of the proposed research arises from (i) establishing NMR-base profiling of IVF media as a tool to non-invasively predict embryo implantation success to reduce the likelihood of multiple IVF pregnancies, and (ii) gaining new insights into the biology of early embryo development. PUBLIC HEALTH RELEVANCE: Treatment with in vitro fertilization (IVF) has resulted in a significant increase in the number of multiple births, significantly compromising both maternal and perinatal health, and increasing health care costs to society. It is imperative to develop non-invasive tools which will allow the selection and transfer of a single embryo in each IVF cycle. We propose a pilot study to characterize the metabolic profile of implantable embryos from conditioned culture media using proton nuclear magnetic resonance spectroscopy (1H-NMR).